Eye movements are used day-to-day to acquire visual information. Vision is also used for postural control. There are growing indications eye movements can affect postural control. However, this has not been investigated in older adults, which is surprising given the high incidence of falls in older populations. The present thesis aims to address this. The first experimental chapter explores the effects of eye movements on balance during standing in young and older adults. The findings show decreased stability during smooth pursuits, whereas saccades maintained stability to that when fixating a static target. The older adults matched the younger groups performance throughout. The second experimental chapter explores the effects of smooth pursuits and saccades on balance during locomotion in young and older adults. Smooth pursuits were shown to decrease stability, whilst saccades maintained stability compared to fixating a static target. The effects of the eye movements were similar in the older adults. However, the elders exhibited lower baseline stability. The third experimental chapter explores the effects of tracking a real-world stimulus (another person known as `pedestrian') on balance control during locomotion. The pedestrian could be standing still or walking. Fixating the stationary and the walking pedestrian decreased stability similarly when compared to free gaze when the pedestrian was not present. To determine whether these results were transferable to natural gaze rather than instructed gaze, the fourth experimental chapter explores free gaze patterns in a similar real-world environment. Both the young and older adults typically fixated the pedestrian when he was standing still and walking, but began to ignore him once he had walked away from their direction heading. Therefore, experiment 3 behaviour was transferable to natural gaze patterns. The older adults also adopted a more cautious approach by fixating regions on the ground initially, and for longer, before looking to their direction heading.

O Simulador de Paciente Humano (Human Patient Simulator - HPS) é um recurso científico e tecnológico de Engenharia Biomédica com soluções de hardware e software que simulam a fisiologia humana. O objetivo é avaliar um caso de uso da simulação clínica com a aplicação do HPS no procedimento de mensuração da pressão arterial (PA) com esfigmomanômetro aneróide e estetoscópio. A pesquisa foi aprovada pelo comitê de ética no parecer 1.544.404/2016. A parte prática do trabalho foi realizada no Laboratório de Simulação de UTI do IFRR. A pesquisa envolveu trinta e três voluntários, estudantes do Curso Técnico em Enfermagem, divididos aleatoriamente em dois grupos. A metodologia proposta utilizou uma pesquisa exploratório-descritiva para extrair dados quantitativos, qualitativos e estatísticos para comparar os resultados de mensuração da PA do Grupo Experimental (GE) com o Grupo Controle (GC). A pesquisa foi desenvolvida em duas partes. Na parte I, um questionário pré-teste foi aplicado inicialmente, experimento de mensuração da PA seguindo o procedimento do respectivo grupo; e o questionário pós-teste. Na parte II, os grupos foram invertidos e submetidos às mesmas etapas da parte I. Coletou-se um total de 599 PAs mensuradas pelos voluntários, sendo 528 PAs do GE e 71 PAs do GC. Com estas PAs foi realizado o teste t de Student ou teste de Wilcoxon com 95% do intervalo de confiança, a depender do teste de linearidade, para comprovar as hipóteses (nula ou alternativa) individualmente e para análise em grupo utiliza o ANOVA ou teste de Kruskal-Wallis possibilitando distinguir quais os voluntários que necessitam de mais treinamento e o respectivo nível da PA. No pós-teste, ao utilizar a escala Likert, os voluntários da pesquisa do GE concordaram em 54% que a metodologia utilizando o HPS fortalece um conhecimento para o desenvolvimento de habilidades clínicas, assim como ser capazes de distinguir sinais normais dos anormais que são propícios para a aprendizagem. Comparando estatisticamente ambos os grupos, o GC destaca que: 53% sabem medir a PA, 13% não sabem medir a PA e 35% das amostras estão inválidas, enquanto o GE apresenta 64% dos voluntários sabem medir a PA, 22% não sabem medir a PA e 14% das amostras estão inválidas. Ademais, os voluntários apresentaram eficácia quando utilizam o HPS. / The Human Patient Simulator (Human Patient Simulator - HPS) is a scientific and technological resource of Biomedical Engineering with hardware and software solutions that simulate human physiology. The objective of this study was to evaluate a case of clinical simulation using an HPS to perform blood pressure (BP) measurement using an aneroid sphygmomanometer and a stethoscope. The research was approved by the ethics committee in the legal opinion 1544.404 / 2016.The practical part of this work was carried out in the ICU Simulation Laboratory of the IFRR. The research involved thirty-three volunteers, who were students of the Nursing Technical Course, randomly divided into two groups. The proposed methodology made use of an exploratory-descriptive research to obtain quantitative, qualitative and statistical data to compare the results of the BP measurement of the Experimental Group (EG) with the Control Group (CG). The research was developed in two parts. In part I, a pre-test questionnaire was initially applied, an experiment of BP measurement following the respective group’s procedure; and a post-test questionnaire. In part II, the groups were inverted and submitted to the same phases of part I. A total of 599 PAs were measured by volunteers, being 528 PAs from the GE and 71 PAs from the GC. With the BP readings, either the Student's t or the Wilcoxon test was performed with a 95% confidence interval, depending on the linearity test, to test the hypotheses (null or alternative) individually, and for the group analysis it was used either the ANOVA or Kruskal's test -Wallis making it possible to distinguish which volunteers need more training and their respective AP level. In the post-test, when the Likert scale was used, there was a 54% agreement among the EG research volunteers that the methodology using the HPS strengthens the knowledge for the development of clinical skills, as well as being able to distinguish normal from abnormal signals that are conducive to learning. Statistically comparing both groups,in the CG: 53% know BP measurement, 13% do not know BP measurement and 35% of the samples are invalid, while in the EG: 64% of the volunteers know BP measurement, 22% do not know how to measure BP and 14% of the samples are invalid. In addition, the volunteers show effectiveness when using the HPS.

The thesis deals with car cabin environment and thermal comfort inside. A car cabin heat load model was developed in Dymola/Modelica to investigate influence of ambient environmental parameters. The model was validated on the data set of eight test cases measured in a climatic chamber and in a real traffic. The main objective of the thesis was to develop a human thermal comfort model suitable for non-homogenous environments and for a car cabin environment especially. The Coupled model of human physiology and thermal comfort was developed in Dymola/Modelica. The model allows predicting an overall human thermal comfort from local boundary conditions representing ambient and personal factors. The model was validated by 16 test cases taken from experiments in literature. Moreover three test cases were created in Theseus-FE to consider an asymmetrical heat load from Sun rays inside a car cabin. Prediction of the Coupled model was compared with Fiala model and experimental data. The Coupled model predicted mean skin temperature for moderate activities in neutral and warm environment well. In cold environment a predicted core temperature was very affected by ambient temperature and during high activity exercises, the predicted mean skin temperature was too high.

The pathogenesis of oral submucous fibrosis (OSMF) is due to a complex interplay between the production and degradation of extracellular matrix (ECM) protein components. In tissue fibrosis, there is a net accumulation of collagen as a result of an imbalance between enhanced production, deposition and impaired degradation of ECM components. OSMF is a chronic inflammatory condition of the oral cavity and regulation of a number of pro-inflammatory and fibrogenic cytokines such as interleukine-1, -6 and -8 isoforms, TGF-β, PDGF, bFGF, IFN-γ and TNF-α has been reported in OSMF tissues. The expression of these growth factors has a bearing on the epithelial changes as well as proliferation and differentiation of oral fibroblasts into ECM protein producing myofibroblast cells. One key modulator of fibrosis in several organs has been TGF-β. Overproduction of TGF-β mRNA and protein has been reported in several fibrotic disorders including that of skin, lungs, liver, kidney and heart. This is mainly due to stimulation of ECM genes by TGF-β. Although there have been few reports suggesting the over production of TGF-β in OSMF tissues, the specific isoforms involved or the mechanisms are poorly understood. Areca nut components, especially arecoline have been implicated in the pathophysiology of OSMF. Few reports indicate the involvement of arecoline in the regulation of collagen production and activity of collagenases and their inhibitors in oral fibroblast cells. Moreover, the alkaloid is involved in initiating epithelial inflammation by inducing COX-2, prostaglandin E2, IL-1α, IL-6 and IL-8 in KB oral carcinoma cells and oral fibroblast cells. These and other reports strongly suggest that changes in gene expression mediated by Arecoline may be central to the progression of OSMF. Not much is known about arecoline-mediated cellular signaling events except for few recent reports that suggest the activation of MAPK pathways. In neuronal and colonic smooth muscle cells of mouse, rat and rabbit, the actions of Arecoline have been reported to be through the activation of muscarinic acetylcholine receptors. Direct binding of arecoline to human M1, 2 and 3 muscarinic receptor isoforms have been shown in brain tissues. Stimulation of these receptors alters the intracellular levels of Ca+2 and cAMP, which are important second messengers. The cholinergic potential of arecoline may be important for their roles in arecoline-mediated signaling events. The expression of muscarinic acetylcholine receptors has been reported in several cell types besides neuronal and excitatory cells. Although several gene expression changes have been reported following Arecoline treatment of a variety of cells, the mechanism of such regulations is not established. Hence in order to understand the role of arecoline in OSMF disease process, we undertook studies that provide insights into arecoline action in epithelial and fibroblast cells and possible molecular mechanisms. The objectives are to study: 1. The role of arecoline in cellular proliferation, cell-cycle regulation and apoptosis in human normal keratinocytes. 2. Mechanism of regulation of gene expression by arecoline in normal keratinocytes. 3. Mechanism of regulation of gene expression by arecoline in human normal oral fibroblasts. In order to achieve the above objectives, a human keratinocyte cell line, HaCaT and an oral periodontal fibroblast cell line (PDC) were utilized. The cells were treated with arecoline and a variety of assays including RT-PCR analysis of mRNA of several genes, phosphorylation status of MAPK pathway intermediates, cell cycle analysis and other cellular and molecular methods have been employed. Following arecoline treatment, there is induction of oxidative stress, growth arrest and epithelial cell death. Since actions of TGF-β are central to most fibrotic disorders and arecoline has been implicated in OSMF, it is hypothesized that arecoline may influence fibrosis via TGF-β pathway. Towards this, several TGF-β target genes that may have a possible role in fibrosis have been studied in arecoline treated epithelial and fibroblast cells. Since arecoline mediated oxidative stress has been reported, the regulation of genes that are involved in stress response pathway have been studied for induction by arecoline in epithelial cells. The results presented in this thesis suggest the up regulation of oxidative stress-responsive genes in HaCaT cells including HOX-1, FTL, G6PD, GCLC and GRD in HaCaT cells. Oxidative stress is a major inducer of inflammatory response in the epithelial tissues. The expression of IL-1α, an important inflammatory cytokine is induced by arecoline in HaCaT cells in response to oxidative stress via the activation of p38 MAPK pathway. Interestingly, activation of MAPK pathways by arecoline is involved in the regulation of common target genes of arecoline and TGF-β and also in the induction of TGF-β−responsive promoter reporter construct, p3TP-lux activity in HaCaT cells. Due to the involvement of TGF-β in fibrosis, regulation of TGF-β pathway genes by arecoline has been studied both in HaCaT and PDC cells. In HaCaT cells, arecoline induces the expression of TGF-β2 mRNA while TβRII expression is down regulated. The expression of the rest of TGF-β/SMAD pathway genes including TGF-β1, β3, TβRI, SMAD1, 2, 3, 4 and 7 are not affected by arecoline in HaCaT cells. Over expression of TGF-β2 is also observed in most of the OSMF tissues compared to normal oral tissues. However, in normal oral fibroblast cells, we observed that the TGF-β/SMAD pathway genes are not regulated by arecoline. These results suggest the possible involvement of arecoline-mediated induction of TGF-β2 in epithelial cells in OSMF disease development. We investigated the signaling pathways involved in the regulation of TGF-β2 and found that stimulation of M3 muscarinic receptor by arecoline leads to the induction of TGF-β2 expression in HaCaT cells via PKC pathway. TGM-2 is an important TGF-β target gene involved in the cross linking of ECM proteins. Arecoline-mediated induction of TGM2 mRNA and transglutaminase activity are observed in oral fibroblast cells, PDC. The induction of TGM-2 was found to be independent of oxidative stress and TGF-β, but dependent on muscarinic acid receptor activation by arecoline and involves cytosolic cAMP. When tested in OSMF tissues, there was an increased expression of TGF-β2, TSP1 and TGM2 as compared to normal tissues suggesting a possible role of these genes in arecoline-mediated progression of OSMF. Interleukin-8 (IL-8), which is involved in inflammation has been reported to be regulated by TGF-β in a cell type specific manner. In several cell types including human endometrial stromal cells, LnCaP (prostate cancer cells), human retinal pigment epithelial cells and rat lung alveolar epithelial (LM5) cells etc., TGF-β up regulates the expression of IL-8 mRNA. Arecoline was found to down regulate IL-8 expression in PDC cells as measured by RT-PCR. Interestingly, the presence of serum along with arecoline induces the expression of IL-8 in PDC cells suggesting the modulation of arecoline-mediated gene regulation by a serum activated signaling pathway. Intriguingly, arecoline treatment led to down regulation of collagens in PDC cells. However, collagen genes are induced in PDC cells in the presence of HaCaT spent medium by arecoline suggesting a role for factor(s) secreted by epithelial cells in the regulation of collagen genes by arecoline. This factor could be an isoform of TGF-β as shown by blocking the induction of collagens by the TGF-β inhibitor, βLAP. Taken together, all these results indicate the ability of arecoline to cause fibrosis in a tissue environment where both epithelial and fibroblasts respond to arecoline and mutually contribute to the disease manifestation. Major conclusions from this study includes, 1] cell death in epithelial cells due to oxidative stress following arecoline treatment, 2] regulation of gene expression by arecoline involves MAPK, PKC pathways, 3] muscarinic acid receptor and oxidative stress are also important for regulation gene expression by arecoline. The most important inference from this study is the possible paracrine influence of TGF-β isoforms secreted by epithelial cells on the oral fibroblasts in determining the progression of OSMF. In summary, in this thesis, an attempt has been made to study the molecular mechanisms and role of arecoline, an alkaloid in conferring gene expression changes that may lead to the initiation and progression of oral sub mucous fibrosis.

A cellular pre-mRNA undergoes various post-transcriptional processing events, including capping, splicing and polyadenylation prior to nuclear export. Splicing is particularly important for mRNA nuclear export as two distinct multi-protein complexes, known as human TREX (hTREX) and the exon-junction complex (EJC), are recruited to the mRNA in a splicing-dependent manner. In contrast, a number of Kaposi's sarcoma-associated herpesvirus (KSHV) lytic mRNAs lack introns and are exported by the virus-encoded ORF57 protein. Herein we show that ORF57 binds to intronless viral mRNAs and functions to recruit the complete hTREX complex, but not the EJC, in order assemble an export component viral ribonucleoprotein particle (vRNP). The formation of this vRNP is mediated by a direct interaction between ORF57 and the hTREX export adapter protein, Aly. Aly in turn interacts directly with the DEAD-box protein UAP56, which functions as a bridge to recruit the remaining hTREX proteins to the complex. Moreover, we show that a point mutation in ORF57 which disrupts the ORF57-Aly interaction leads to a failure in the ORF57-mediated recruitment of the entire hTREX complex to the intronless viral mRNA and inhibits the mRNAs subsequent nuclear export and virus replication. Furthermore, we have utilised a trans-dominant Aly mutant to prevent the assembly of the complete ORF57-hTREX complex; this results in a vRNP consisting of viral mRNA bound to ORF57, Aly and the nuclear export factor, TAP. Strikingly, although both the export adapter Aly and the export factor TAP were present on the viral mRNP, a dramatic decrease in intronless viral mRNA export and virus replication was observed in the absence of the remaining hTREX components (UAP56 and hTHO-complex). Together, these data provide the first direct evidence that the complete hTREX complex is essential for the export of KSHV intronless mRNAs and infectious virus production.

Acompanha: Uma sequência didática para o ensino de anatomia e fisiologia humana / Este estudo apresenta uma proposta de sequência didática que insere vídeos como mediadores no processo de ensino e aprendizagem de Anatomia e Fisiologia Humana para o oitavo ano do ensino fundamental. A investigação foi realizada em uma escola estadual de um dos municípios, da região metropolitana de Curitiba e busca responder a seguinte questão: Como diferentes categorias de vídeos, inseridos em uma sequência didática contribuem para o processo de ensino aprendizagem? Para responder tal questão a pesquisa foi embasada nas teorias de aprendizagem de Vygotsky.A falta de pesquisa na área de ensino de Anatomia e Fisiologia Humana e o grande interesse dos alunos em notícias e questões da área médica e de saúde pública foi a maior motivação da escolha do tema. A escolha do uso de vídeos como ferramenta para a sequência didática se deu pela conclusão de que através destes recursos o ensino pode se tornar mais atrativo e prazeroso. Os vídeos foram classificados conforme Bartolomé como “Lição”, “Monoconceitual”, e “Impacto”, e como documento da coleta de dados foi utilizado dois relatórios que foram analisados de forma comparativa, o que permitiu qualificar os ganhos em relação a aprendizagem dos conteúdos de Anatomia e Fisiologia Humana mostrando principalmente, a importância de inserir vídeos de diferentes categorias, onde cada tipologia aborda o conteúdo com uma perspectiva diferenciada dentro das várias metodologias de ensino, podendo ter a intervenção do professor e aluno, principalmente do vídeo classificado como sendo de impacto. / This study presents a proposal for a didactic sequence that inserts videos as mediators in the teaching and learning process of Anatomy and Human Physiology for the eighth year of elementary school. The research was carried out at a state school in one of the municipalities of the metropolitan region of Curitiba and seeks to answer the following question: How do different categories of videos, inserted in a didactic sequence, contribute to the learning process? To answer such a question the research was based on Vygotsky's theories of learning. The lack of research in the field of Anatomy and Human Physiology teaching and the great interest of the students in medical and public health news and issues was the major motivation for choosing the topic. The choice of videos as a tool for the didactic sequence was based on the conclusion that through these resources teaching can become more attractive and enjoyable. The videos were classified according to Bartolomé as "Lesson", "Monoconceitual", and "Impacto", and as a data collection document two reports were used that were analyzed in a comparative way, which allowed to qualify the gains in relation to the learning of the contents Of Anatomy and Human Physiology showing mainly the importance of inserting videos of different categories,where each typology approaches the content with a different perspective within the various teaching methodologies, being able to have the intervention of the teacher.

Acompanha: Uma sequência didática para o ensino de anatomia e fisiologia humana / Este estudo apresenta uma proposta de sequência didática que insere vídeos como mediadores no processo de ensino e aprendizagem de Anatomia e Fisiologia Humana para o oitavo ano do ensino fundamental. A investigação foi realizada em uma escola estadual de um dos municípios, da região metropolitana de Curitiba e busca responder a seguinte questão: Como diferentes categorias de vídeos, inseridos em uma sequência didática contribuem para o processo de ensino aprendizagem? Para responder tal questão a pesquisa foi embasada nas teorias de aprendizagem de Vygotsky.A falta de pesquisa na área de ensino de Anatomia e Fisiologia Humana e o grande interesse dos alunos em notícias e questões da área médica e de saúde pública foi a maior motivação da escolha do tema. A escolha do uso de vídeos como ferramenta para a sequência didática se deu pela conclusão de que através destes recursos o ensino pode se tornar mais atrativo e prazeroso. Os vídeos foram classificados conforme Bartolomé como “Lição”, “Monoconceitual”, e “Impacto”, e como documento da coleta de dados foi utilizado dois relatórios que foram analisados de forma comparativa, o que permitiu qualificar os ganhos em relação a aprendizagem dos conteúdos de Anatomia e Fisiologia Humana mostrando principalmente, a importância de inserir vídeos de diferentes categorias, onde cada tipologia aborda o conteúdo com uma perspectiva diferenciada dentro das várias metodologias de ensino, podendo ter a intervenção do professor e aluno, principalmente do vídeo classificado como sendo de impacto. / This study presents a proposal for a didactic sequence that inserts videos as mediators in the teaching and learning process of Anatomy and Human Physiology for the eighth year of elementary school. The research was carried out at a state school in one of the municipalities of the metropolitan region of Curitiba and seeks to answer the following question: How do different categories of videos, inserted in a didactic sequence, contribute to the learning process? To answer such a question the research was based on Vygotsky's theories of learning. The lack of research in the field of Anatomy and Human Physiology teaching and the great interest of the students in medical and public health news and issues was the major motivation for choosing the topic. The choice of videos as a tool for the didactic sequence was based on the conclusion that through these resources teaching can become more attractive and enjoyable. The videos were classified according to Bartolomé as "Lesson", "Monoconceitual", and "Impacto", and as a data collection document two reports were used that were analyzed in a comparative way, which allowed to qualify the gains in relation to the learning of the contents Of Anatomy and Human Physiology showing mainly the importance of inserting videos of different categories,where each typology approaches the content with a different perspective within the various teaching methodologies, being able to have the intervention of the teacher.

Indiana University-Purdue University Indianapolis (IUPUI) / Atherosclerotic lesions of human beings are common diagnosed in regions of arte- rial branching and curvature. The prevalence of atherosclerosis is usually associated with hardening and ballooning of aortic wall surfaces because of narrowing of flow path by the deposition of fatty materials, platelets and influx of plasma through in- timal wall of Aorta. High Wall Shear Stress (WSS) is proved to be the main cause behind all these aortic diseases by physicians and researchers. Due to the fact that the atherosclerotic regions are associated with complex blood flow patterns, it has believed that hemodynamics and fluid-structure interaction play important roles in regulating atherogenesis. As one of the most complex flow situations found in cardio- vascular system due to the strong curvature effects, irregular geometry, tapering and branching, and twisting, theoretical prediction and in vivo quantitative experimental data regarding to the complex blood flow dynamics are substantial paucity. In recent years, computational fluid dynamics (CFD) has emerged as a popular research tool to study the characteristics of aortic flow and aim to enhance the understanding of the underlying physics behind arteriosclerosis. In this research, we study the hemo- dynamics and flow-vessel interaction in patient specific normal (healthy) and dilated (diseased) aortas using Ansys-Fluent and Ansys-Workbench. The computation con- sists of three parts: segmentation of arterial geometry for the CFD simulation from computed tomography (CT) scanning data using MIMICS; finite volume simulation of hemodynamics of steady and pulsatile flow using Ansys-Fluent; an attempt to perform the Fluid Structure Simulation of the normal aorta using Ansys-Workbench. Instead of neglecting the branching or smoothing out the wall for simplification as a lot of similar computation in literature, we use the exact aortic geometry. Segmen- tation from real time CT images from two patients, one young and another old to represent healthy and diseased aorta respectively, is on MIMICS. The MIMICS seg- mentation operation includes: first cropping the required part of aorta from CT dicom data of the whole chest, masking of the aorta from coronal, axial and saggital views of the same to extract the exact 3D geometry of the aorta. Next step was to perform surface improvement using MIMICS 3-matic module to repair for holes, noise shells and overlapping triangles to create a good quality surface of the geometry. A hexahe- dral volume mesh was created in T-Grid. Since T-grid cannot recognize the geometry format created by MIMICS 3-matic; the required step geometry file was created in Pro-Engineer. After the meshing operation is performed, the mesh is exported to Ansys Fluent to perform the required fluid simulation imposing adequate boundary conditions accordingly. Two types of study are performed for hemodynamics. First is a steady flow driven by specified parabolic velocity at inlet. We captured the flow feature such as skewness of velocity around the aortic arch regions and vortices pairs, which are in good agreement with open data in literature. Second is a pulsatile flow. Two pulsatile velocity profiles are imposed at the inlet of healthy and diseased aorta respectively. The pulsatile analysis was accomplished for peak systolic, mid systolic and diastolic phase of the entire cardiac cycle. During peak systole and mid-systole, high WSS was found at the aortic branch roots and arch regions and diastole resulted in flow reversals and low WSS values due to small aortic inflow. In brief, areas of sudden geometry change, i.e. the branch roots and irregular surfaces of the geom- etry experience more WSS. Also it was found that dilated aorta has more sporadic nature of WSS in different regions than normal aorta which displays a more uniform WSS distribution all over the aorta surface. Fluid-Structure Interaction simulation is performed on Ansys-WorkBench through the coupling of fluid dynamics and solid mechanics. Focus is on the maximum displacement and equivalent stress to find out the future failure regions for the peak velocity of the cardiac cycle.